Single-cell RNA sequencing (scRNA-seq) data often encountered technical artifacts called "doublets" which are two cells that are sequenced under the same cellular barcode.
Doublets formed from different cell types or states are called heterotypic and homotypic otherwise. These factors constrain cell throughput and may result in misleading biological interpretations.
DoubletFinder (McGinnis, Murrow, and Gartner 2019) is one of the methods proposed for doublet detection. In this notebook, we will illustrate an example workflow of DoubletFinder. We use a 10x Genomics dataset which captures peripheral blood mononuclear cells (PBMCs) from a healthy donor stained with a panel of 31 TotalSeq™-B antibodies (BioLegend).
Build single-cell trajectories with the software that introduced **pseudotime**. Find out about cell fate decisions and the genes regulated as they're made.
Group and classify your cells based on gene expression. Identify new cell types and states and the genes that distinguish them.
Find genes that vary between cell types and states, over trajectories, or in response to perturbations using statistically robust, flexible differential analysis.
In development, disease, and throughout life, cells transition from one state to another. Monocle introduced the concept of **pseudotime**, which is a measure of how far a cell has moved through biological progress.
Many researchers are using single-cell RNA-Seq to discover new cell types. Monocle 3 can help you purify them or characterize them further by identifying key marker genes that you can use in follow-up experiments such as immunofluorescence or flow sorting.
**Single-cell trajectory analysis** shows how cells choose between one of several possible end states. The new reconstruction algorithms introduced in Monocle 3 can robustly reveal branching trajectories, along with the genes that cells use to navigate these decisions.
InferCNV is used to explore tumor single cell RNA-Seq data to identify evidence for somatic large-scale chromosomal copy number alterations, such as gains or deletions of entire chromosomes or large segments of chromosomes. This is done by exploring expression intensity of genes across positions of tumor genome in comparison to a set of reference 'normal' cells. A heatmap is generated illustrating the relative expression intensities across each chromosome, and it often becomes readily apparent as to which regions of the tumor genome are over-abundant or less-abundant as compared to that of normal cells.
**Infercnvpy** is a scalable python library to infer copy number variation (CNV) events from single cell transcriptomics data. It is heavliy inspired by InferCNV, but plays nicely with scanpy and is much more scalable.
In this notebook, we present COMMOT (COMMunication analysis by Optimal Transport) to infer cell-cell communication (CCC) in spatial transcriptomic, a package that infers CCC by simultaneously considering numerous ligand–receptor pairs for either spatial transcriptomic data or spatially annotated scRNA-seq data equipped with spatial distances between cells estimated from paired spatial imaging data.
A collective optimal transport method is developed to handle complex molecular interactions and spatial constraints. Furthermore, we introduce downstream analysis tools to infer spatial signaling directionality and genes regulated by signaling using machine learning models.
Integration of single-cell RNA sequencing (scRNA-seq) data from multiple experiments, laboratories, and technologies can uncover biological insights, but current methods for scRNA-seq data integration are limited by a requirement for datasets to derive from functionally similar cells. We present Scanorama, an algorithm that identifies and merges the shared cell types among all pairs of datasets and accurately integrates heterogeneous collections of scRNA-seq data.
Scanorama enables batch-correction and integration of heterogeneous scRNA-seq datasets, which is described in the paper "Efficient integration of heterogeneous single-cell transcriptomes using Scanorama" by Brian Hie, Bryan Bryson, and Bonnie Berger.
Scanorama is designed to be used in scRNA-seq pipelines downstream of noise-reduction methods, including those for imputation and highly-variable gene filtering. The results from Scanorama integration and batch correction can then be used as input to other tools for scRNA-seq clustering, visualization, and analysis.